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LC MSMS Allergen Screening Method for EU1169/2011

In collaboration with the Wageningen University BU Bioscience, Nutrilab has developed a method to measure more than 21 allergens in 1 LC-MSMS run. The big advantage of this method is that it significantly reduces the cost price of simultaneously analysing several allergens in a sample. The method has additional multiple benefits in comparison to the traditional ELISA and PCR techniques.

Measurement and Method

The method comprises the analysis of all proteins based on allergens listed in EU1169/2011, and a few allergens for non-European markets. For the sake of clarity, these are the allergens [based on the foods in which they occur:

Allergens

Allergens from nuts

Gluten

Mustard

Almonds

Shellfish

Sesame

Hazelnuts

Egg

Lupine

Pistachios

Fish

Mollusks

Cashew nuts

Groundnuts (peanuts)

Buckwheat

Walnuts

Soy

Fenugreek

Macadamia nuts

Milk

Brazil nuts

Celery

Pecan nuts

Nutrilab’s LC-MSMS method for allergen identification uses three or more different protein fragments (peptides) present in the allergen in question. For each peptide, at least 2 or more transitions are used for the MS (mass spectrometer). At the same time the mass ratios between the various allergenic proteins in a sample is used for the assessment.  This becomes clearer when using peanuts as an example. Peanuts contain many different proteins. The primary allergens in peanuts are mainly AraH1, AraH2, AraH3, AraH4, AraH5 and AraH6. In peanuts, these proteins occur in fixed mass ratios. By observing the occurrence of these proteins AND their ratios, a very high degree of reliability and selectivity is achieved.

With the Nutrilab LC-MSMS method, the screening has an LOD for all allergens in the range of 0.1 - 4 ppm depending on the allergen.

Advantages compared to ELISA and PCR

The PCR method searches for the presence of DNA of associated with a particular protein. In most cases the particular protein is present as well as the associated DNA. The DNA is used to identify the species involved, such as celery or mustard, but not the specific allergen. Furthermore, it is difficult to translate the amount of DNA found to the product’s protein fraction. This introduces a large amount of spread in the results and thus makes it very difficult to compare the results obtained from PCR and ELISA.

In the ELISA method the different producers make different antibodies to target the product’s proteins. It is also not known whether antibodies target proteins from processed food or a pure protein. Consequently, the ELISA results generated by different manufacturers are often not at all comparable. There are manufacturers of ELISA kits and rapid allergen tests who can achieve very low LODs but with a totally unreliable method. The LC-MSMS method offers many more advantages compared to ELISA because proteins and protein fragments are examined, regardless of whether a product is pure or processed.

For more information, or to submit samples, please contact: Nutrilab

Allergenen

Increasing numbers of consumers are now aware that food can cause hypersensitivity reactions and allergies. In response, the food allergy research programme is being extended. Furthermore, regulations have been introduced making it compulsory to the presence of allergens on product labels.

Allergens are proteins that can cause allergic reactions. The European Union has determined that food allergens that cause the most allergic reactions should be listed on the label.

At Nutrilab the allergen testing is being carried out with modern analytical techniques, ELISA and PCR.

- Almond
- Beta-lactoglobulin
- Casein
- Celery
- Egg
- Fish
- Gluten-containing grains like wheat, rye, barley, oats, spellt, kamut
- Histamine
- Lactose
- Lupine
- Milk
- Molluscs
- Mustard
- Nuts such as almond, hazelnut, walnut, cashew, pecan, Brazil nut, pistachio and macadamia nuts
- Peanut
- Sesame
- Shellfish
- Soy
- Sulphite